KR20180082971A - Stable Liquid Formulation - Google Patents

Abstract

According to the present invention, a stable liquid preparation comprises: an antibody or an antigen binding site therefor; an acetate buffer; glycine; and a surfactant, without containing at least one of sugar, sugar alcohol, and a metal salt. The stable liquid preparation is stable even when containing high amount of antibodies, is excellent in terms of osmotic concentration and viscosity, and can be administered subcutaneously.

Description

[0001] Stable Liquid Formulation [

The present invention relates to a stable liquid formulation.

Protein preparations, especially intravenous injectable antibody preparations have been used for a considerable period of time. Proteins, particularly antibodies, form aggregates and / or dimers and tend to be fragmented or denatured. If the formulation containing it is injected intravenously, it can cause serious side effects, such as anaphylactic shock. Numerous methods have been attempted to prevent such aggregation and fragmentation and to improve its stability. For example, antibodies for intravenous use are often lyophilized to improve stability during storage, but such preparations must be reconstituted with a diluent prior to use. The reconstitution step is inconvenient, time-consuming and increases the possibility of contamination of the product.

Various liquid formulations are known as conventional formulations containing antibodies. U.S. Patent No. 8932591 discloses a stable liquid pharmaceutical formulation comprising an anti-hTNFa antibody, a polyol, a surfactant, and a buffer system (citrate and phosphate). However, this preparation may be restricted in the number of administration times and the administration period due to the low antibody content of about 50 mg / mL, and there is still a need for improvement in stability.

U.S. Patent Publication No. 2005-0260204 discloses an antibody preparation comprising an antibody, histidine, polyol and / or NaCl. However, in the case of this liquid preparation, problems such as precipitation and gelatinization including NaCl as an isotonic agent may occur, and there is still a need for improvement in terms of stability.

Korean Patent Laid-Open Publication No. 10-2014-0134689 discloses an effective amount of a stabilizer selected from a buffer system (succinate), a surfactant, an isotonizing agent (NaCl or KCl), and an amino acid (arginine) and a cyclodextrin, Lt; / RTI > antibody or antigen-binding portion thereof. However, in the case of this liquid preparation, problems such as precipitation and gelatinization may occur including NaCl or KCl as an isotonic agent, and the number of administration and the period of administration may be limited due to the low antibody content of about 50 mg / mL .

Thus, there is a need for a stable liquid formulation that can overcome the problems of the conventional liquid formulation and comprise an antibody.

It is an object of the present invention to provide a stable liquid preparation.

Another object of the present invention is to provide a stable liquid formulation even when the antibody is contained in a high content.

Another object of the present invention is to provide a stable liquid preparation excellent in osmosis and viscosity.

It is still another object of the present invention to provide a stable liquid preparation capable of subcutaneous administration.

In one embodiment of the invention, a stable liquid preparation comprises an antibody or antigen binding portion thereof; Acetate buffer; Glycine; And a surfactant, and may not contain at least one of a sugar, a sugar alcohol, and a metal salt.

In one embodiment of the invention, the (A) antibody may comprise a monoclonal antibody.

In one embodiment of the invention, (A) the antibody may comprise a fully human antibody.

In one embodiment of the invention, (A) the antibody may comprise an antibody that binds to TNF- [alpha].

In one embodiment of the invention, (A) the antibody may comprise one or more of infliximag, adalimumab, sertolium febulol, and golimumim.

In one embodiment of the invention, the antibody (A) comprises a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3 Light chain variable region; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6.

In one embodiment of the invention, the antibody (A) comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7; And a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

In one embodiment of the invention, the antibody (A) comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9; And a heavy chain comprising the amino acid sequence of SEQ ID NO: 10.

In one embodiment of the invention, the concentration of (A) antibody may be between 50 and 150 mg / mL.

In one embodiment of the invention, (B) the acetate buffer solution may comprise an acetate salt.

In one embodiment of the invention, the acetic acid salt content may be between 1 and 30 mM.

In one embodiment of the invention, the stable liquid formulation may not contain one or more of histidine, citrate, phosphate, maleate, tartrate, and succinate.

In one embodiment of the invention, the concentration of (C) glycine may be between 100 and 300 mM.

In one embodiment of the invention, the stable liquid preparation is a liquid formulation comprising one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, ≪ / RTI > and valine.

In one embodiment of the present invention, (D) the surfactant may comprise polysorbate, poloxamer, or mixtures thereof.

In one embodiment of the present invention, (D) the surfactant may comprise at least one of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.

In one embodiment of the present invention, (D) the surfactant may comprise polysorbate 80.

In one embodiment of the invention, the concentration of (D) surfactant may be from 0.01 to 1% (w / v).

In one embodiment of the invention, the pH may be between 4.5 and 5.5.

In one embodiment of the invention, the osmolality concentration may be from 200 to 400 mmol / kg.

In one embodiment of the invention, the stable liquid formulation may not contain a preservative, a chelating agent or a mixture thereof.

In one embodiment of the invention, the stable liquid preparation comprises 50 to 150 mg / mL of the antibody or antigen-binding portion thereof; Acetic acid salt buffer containing 1 to 30 mM acetate; 100 to 300 mM glycine; And 0.01 to 1% (w / v) of a surfactant, and may not contain at least one of a sugar, a sugar alcohol, and a metal salt.

In one embodiment of the invention, the stable liquid formulation comprises (A) a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: A light chain variable region comprising a light chain variable region; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6, Binding portion 50 to 150 mg / mL; (B) an acetic acid salt buffer containing 1 to 30 mM acetate; (C) glycine 100 to 300 mM; And (D) 0.01 to 1% (w / v) of a surfactant, and may not contain at least one of a sugar, a sugar alcohol, and a metal salt.

In one embodiment of the invention, the stable liquid formulation comprises (A) a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: A light chain variable region comprising a light chain variable region; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6, Binding portion 100 mg / mL; (B) Acetate buffer solution containing 10 mM acetic acid salt; (C) glycine 250 mM; And (D) 0.1% (w / v) of a surfactant, and may not contain at least one of a sugar, a sugar alcohol, and a metal salt.

In one embodiment of the invention, the stable liquid formulation may be for subcutaneous administration.

In another embodiment of the present invention, a pre-filled syringe filled with a stable liquid formulation is provided.

In another embodiment of the present invention, an auto-injector is provided in which a pre-field syringe is contained.

The stable liquid preparation according to the present invention is stable even when it contains a high amount of antibody, is excellent in osmotic concentration and viscosity, and can be administered subcutaneously.

[Stable liquid preparation]

In one embodiment of the invention, a stable liquid preparation comprises an antibody or antigen binding portion thereof; Acetate buffer; Glycine; And a surfactant, and may not contain at least one of a sugar, a sugar alcohol, and a metal salt.

&Quot; Stable " or " Stability "

In the context of the present application, the term " stable " or " stability " means that an antibody according to the present invention substantially retains its physical stability and / or chemical stability and / or biological activity during the manufacturing process and / it means. A variety of analytical techniques for measuring antibody stability are readily available in the art.

The physical stability can be assessed by methods known in the art, including the measurement of sample apparent attenuation of light (absorbance or optical density). This light attenuation measurement is related to the turbidity of the formulation. Further, high molecular weight component content, low molecular weight component content, intact protein amount, and insoluble foreign particle number can be measured with respect to physical stability.

Chemical stability can be assessed, for example, by detecting and quantifying chemically altered forms of the antibody. Chemical stability includes, for example, charge changes that may be assessed by ion exchange chromatography (such as occurs as a result of deamidation or oxidation). For chemical stability, charge modifications (acidic or basic peaks) and the like can be measured.

Biological activity can be assessed by methods known in the art and antigen binding affinity can be determined, for example, by ELISA.

The term " not comprising " in the specification of the present application means not containing any of the components. Also, the term is intended to include those that do not substantially contain the component, i. E., The activity of the antibody, the stability and viscosity of the liquid formulation, such as, for example, 1% (w / v), 0 to 1 ppm (w / v) or 0 to 1 ppb (w / v).

(A) Antibody

An antibody refers to an immunoglobulin molecule consisting of four polypeptide chains in which two heavy chains and two light chains are linked to each other by disulfide bonds. Other naturally occurring antibodies with altered structures, such as camelid antibodies, are also included in this definition. Each heavy chain consists of a heavy chain variable region and a heavy chain constant region. The heavy chain constant region consists of three domains (CH1, CH2 and CH3). Each light chain consists of a light chain variable region and a light chain constant region. The light chain constant region consists of one domain (CL). The heavy chain variable region and the light chain variable region can be further subdivided into a hypervariable region called the complementarity determining region (CDR), which is disposed with a more conserved region called the framework region (FR). Each heavy chain variable region and light chain variable region consists of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

In one embodiment of the invention, the antibody can include a polyclonal antibody, a monoclonal antibody, a recombinant antibody, a single chain antibody, a hybrid antibody, a chimeric antibody, a humanized antibody, a fully human antibody, or a fragment thereof. In one embodiment of the invention, a monoclonal antibody can be included as an antibody. Chimeric human-mouse monoclonal antibodies can be prepared by methods known in the art. For example, in the case of infliximab, it can be prepared by the method described in U.S. Patent No. 6,284,471. Fully human antibodies have been created to reduce the side effects of humanized antibodies or chimeric antibodies, and transgenic mice and phage display have been identified as successful recipes. In one embodiment of the invention, it may comprise a fully human antibody as an antibody. A fully human monoclonal antibody can be prepared by a known method. For example, Adalimumab can be prepared by the method described in U.S. Patent No. 6,090,382.

In one embodiment of the invention, the antibody may comprise an antibody that binds to an epitope of TNF- [alpha] or TNF- [alpha]. Antibodies that bind to an epitope of TNF- [alpha] or TNF- [alpha] may include infliximag, adalimumab, sertolium mappalol, golimamap, or mixtures thereof. In one embodiment of the invention, it may comprise asparagine as an antibody.

In one embodiment of the invention, the antibody comprises a light chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: domain; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6.

In one embodiment of the invention, the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7; And a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

In one embodiment of the invention, the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9; And a heavy chain comprising the amino acid sequence of SEQ ID NO: 10.

In the present invention, the concentration of the antibody may be 50 mg / mL or more. The concentration of the antibody can be freely adjusted within a range that does not substantially adversely affect the stability and viscosity of the stable liquid formulation according to the present invention. In one embodiment of the invention, the concentration of the antibody is in the range of 50 to 150 mg / mL, such as 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 mg / mL. In another embodiment of the invention, the concentration of the antibody is from 55 to 145 mg / mL, from 60 to 140 mg / mL, from 65 to 135 mg / mL, from 70 to 130 mg / mL, from 75 to 125 mg / mg / mL, 85 to 115 mg / mL, 90 to 110 mg / mL or 95 to 105 mg / mL. When the concentration of the antibody is within the above range, the degree of freedom of the administration dose and the administration period can be increased according to the high content of the antibody, and the stability, viscosity and formulation of the preparation can be excellent.

(B) Acetate buffer

The acetate buffer solution may comprise an acetate salt. Examples of the acetic acid salt include, but are not limited to, sodium acetate, zinc acetate, aluminum acetate, ammonium acetate, and potassium acetate. The acetate buffer solution can be prepared by mixing the acetate salt with acetic acid. In one embodiment of the invention, the acetate buffer solution may comprise sodium acetate.

In one embodiment of the invention, the stable liquid formulation may not contain other buffers. In one embodiment of the invention, the stable liquid formulation may not contain histidine, citrate, phosphate, maleate, tartrate, succinate or mixtures thereof. The stability and viscosity of the formulation may be deteriorated when the buffer solution is used in place of or in addition to the acetate buffer solution, and the stability of the preparation may be poor, especially under ultraviolet / harsh conditions, I can feel it.

In one embodiment of the invention, the acetic acid salt content in the acetate buffer can be adjusted freely within a range that does not substantially adversely affect the stability, viscosity and osmolality of the liquid formulation according to the invention. For example, the acetate salt content may be in the range of 0.1 to 45 mM, 1 to 30 mM, 1 to 25 mM, or 5 to 15 mM, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 mM. When the content of the acetate salt is within the above range, the stability of the preparation, osmotic concentration and viscosity can be excellent.

On the other hand, the content of the acetate salt is the content of the acetate salt in the preparation stored in one container (vial or free-field syringe), and in the case of a container intended for distribution or multiple administration, The content may increase several times. Conversely, in the case of small vessels, the content of acetate may be reduced accordingly.

(C) Glycine

In one embodiment of the invention, the stable liquid formulation comprises glycine in an amino acid. Glycine can act as a stabilizer and can contribute to the regulation of physiological osmolality. In one embodiment of the invention, the stable liquid preparation is a liquid formulation comprising one or more of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, ≪ / RTI > and valine. When the above-mentioned other amino acid is contained instead of or in addition to glycine, the solubility may be poor and the liquid formulation may not be possible or the stability may be poor.

In one embodiment of the invention, the (C) glycine content is in the range of 100 to 300 mM, 150 to 300 mM, or 200 to 300 mM, such as 100, 101, 102, 103, 104, 105, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 137, 138, 139, 140, 141, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 297, 298, 299 or 300 mM. When the content of glycine is within this range, it can be excellent in osmotic concentration, stability of the preparation (insoluble foreign particle) and viscosity.

(D) Surfactant

Examples of surfactants include polyoxyethylene sorbitan fatty acid esters (e.g., polysorbates), polyoxyethylene alkyl ethers (e.g., Brij), alkylphenyl polyoxyethylene ethers (e.g., Triton-X) But are not limited to, polyoxyethylene-polyoxypropylene copolymers (e.g., Poloxamer, Pluronic), sodium dodecyl sulfate (SDS), and the like. In one embodiment of the present invention, (D) the surfactant may comprise polysorbate, poloxamer, or mixtures thereof. In one embodiment of the present invention, (D) the surfactant may comprise a polyoxyethylene sorbitan fatty acid ester (polysorbate). In one embodiment of the present invention, (D) the surfactant may comprise at least one of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. In one embodiment of the invention, (D) the surfactant may comprise polysorbate 20, polysorbate 80 or mixtures thereof. In one embodiment of the present invention, (D) the surfactant may comprise polysorbate 80.

In one embodiment of the invention, the concentration of (D) surfactant can be freely adjusted within a range that does not adversely affect the stability and viscosity of the stable liquid formulation according to the present invention. In one embodiment of the invention, the concentration of (D) surfactant is in the range of 0.001 to 5% (w / v), 0.01 to 1% (w / v), 0.02 to 1% (w / v), such as 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5% (w / v). When the concentration of the surfactant (D) is within this range, excellent stability and viscosity can be exhibited.

No or additional ingredients

In one embodiment of the invention, the stable liquid formulation may not contain at least one of a sugar, a sugar alcohol, and a metal salt.

The sugar may not include monosaccharides, disaccharides, oligosaccharides, polysaccharides, or a mixture of two or more thereof. Examples of monosaccharides include, but are not limited to, glucose, fructose, galactose, and the like. Examples of disaccharides include, but are not limited to, sucrose, lactose, maltose, trehalose, and the like. Examples of oligosaccharides include, but are not limited to, fructooligosaccharides, galactooligosaccharides, and mannanoligosaccharides. Examples of polysaccharides include, but are not limited to, starch, glycogen, cellulose, chitin, pectin, and the like.

Examples of sugar alcohols include glycerol, erythritol, traceol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, bolemitol, isomalt, maltitol, lactitol, maltotriol , Maltotetriol, polyglycitol, and the like.

In one embodiment of the invention, the sugar or sugar alcohols may not include sorbitol, mannitol, trehalose, sucrose or a mixture of two or more thereof.

The inclusion of sugars or sugar alcohols increases the viscosity of the liquid formulation, which allows the patient to experience greater pain during subcutaneous injection.

In one embodiment of the present invention, it is possible as a metal salt does not include NaCl, KCl, NaF, KBr, NaBr, Na ₂ SO _4, NaSCN, K ₂ SO ₄ or mixtures thereof. When these compounds are included, a precipitation phenomenon may occur and the preparation may have the shape of gelatin and the stability may be poor.

In one embodiment of the invention, a chelating agent (e.g., EDTA) may not be included. The inclusion of a chelating agent may increase the oxidation rate.

In one embodiment of the invention, it may not contain a preservative. Examples of preservatives include octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl alcohol, benzyl alcohol, alkyl paraben, catechol, resorcinol, cyclohexanol, M-cresol, and the like. Containing a preservative may not help to improve stability.

In one embodiment of the invention, it may further comprise additives known in the art within the scope of substantially not adversely affecting the activity of the antibody, stability and viscosity of the formulation in stable liquid preparations. For example, an aqueous carrier, an antioxidant, or a mixture of two or more thereof. Aqueous carriers are those that are pharmaceutically acceptable (safe and non-toxic when administered to humans) and useful for the preparation of liquid preparations. Examples of aqueous carriers include, but are not limited to, sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), sterile saline solution, Ringer's solution, dextrose, and the like. Antioxidants include, but are not limited to, ascorbic acid.

pH

In one embodiment of the invention, the pH of the stable liquid formulation may be from 4.5 to 5.5, for example 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4 or 5.5. In one embodiment of the invention, the pH can be adjusted using an acetate buffer. In other words, when the acetic acid buffer solution is contained in a predetermined amount, the pH in the above range can be exhibited even without a separate pH adjusting agent. When using buffers containing histidine, citrate, phosphate, maleate, tartarate, succinate or mixtures thereof, it may be difficult to show the pH in the above range. The stability of the antibody may be degraded if it further comprises an acid or a base (for example, sodium hydroxide) as a separate pH adjusting agent.

Osmolality

In one embodiment of the invention, the osmolality of the stable liquid formulation is in the range of 200 to 400 mmol / kg, 250 to 350 mmol / kg, or 270 to 330 mmol / kg, such as 200, 201, 202, 220, 221, 222, 223, 224, 225, 226, 227, 228, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 234, 235, 236, 237, 238, 239, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 385, 386, 387, 388, 389, 382, 383, 384, 385, 386, 374, 375, 376, 377, 378, 379, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399 or 400 mmol / kg. When the osmotic concentration is within the above range, the pain that may occur upon subcutaneous administration can be minimized. In one embodiment of the invention, the osmolality concentration can be controlled using acetate buffer and glycine. In other words, when the acetic acid salt buffer and glycine are contained in a predetermined amount, the osmolality can be expressed in the above range without any additional osmotic pressure regulator. In the case of additionally containing NaCl or the like as a separate osmotic pressure regulating agent, precipitation may occur and the preparation may have the shape of gelatin and the stability may be poor.

Viscosity

In one embodiment of the invention, the viscosity of the stable liquid formulation measured after storage at room temperature (25 캜 3 캜) immediately after formulation or after storage for 6 weeks at 5 캜 3 캜 or 40 캜 2 캜 is 0.5 1.5 to 1.6 cps, such as 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0 cp Lt; / RTI > When the viscosity is within the above range, the pain that may occur upon subcutaneous administration can be minimized, the formulation can be facilitated, the stability of the preparation can be excellently demonstrated, and when applied to a pre-field syringe or an automatic syringe, Can be excellent in terms of a plunger-stopper break loose force or a dynamic glide force.

In one embodiment of the invention, the term " stable " liquid formulation may mean a liquid formulation that satisfies one or more of the following:

Appearance analysis

A liquid formulation in which the clarity of the formulation observed with the naked eye after storage for 6 weeks at a temperature of 5 ° C ± 3 ° C is clear;

A liquid formulation at a temperature of 5 째 C 賊 3 째 C and a clearness of clearness of the preparation observed under visual observation after storage for 6 weeks in an airtight condition;

A liquid formulation in which the clarity of the formulation observed with the naked eye after storage for 6 weeks at a temperature of 40 ° C ± 2 ° C is clear;

A liquid formulation having a temperature of 40 ° C ± 2 ° C, a relative humidity of 75 ± 5%, and a clearness of the visual observation of the formulation after storage for 6 weeks in an airtight condition.

Turbidity

A liquid preparation having an absorbance of 0 to 0.0900 as measured at 350 nm in a spectrophotometer after storage for 6 weeks at a temperature of 5 ° C ± 3 ° C;

A liquid formulation having a temperature of 5 ° C ± 3 ° C and an absorbance of 0 to 0.0900 as measured at 350 nm in a spectrophotometer after storage for 6 weeks in an airtight condition;

A liquid preparation having an absorbance of 0 to 0.1300 as measured at 350 nm in a spectrophotometer after storage for 6 weeks at a temperature of 40 ° C ± 2 ° C;

A liquid formulation having a temperature of 40 ° C ± 2 ° C, a relative humidity of 75 ± 5%, and an absorbance of 0 to 0.1300 as measured at 350 nm in a spectrophotometer after storage for 6 weeks in an airtight condition;

- a liquid preparation having an absorbance of 0 to 0.0900 as measured at 350 nm in a spectrophotometer after storage for 3 weeks at a temperature of 45 ° C ± 2 ° C.

High molecular weight components (peaks with a retention time ahead of the main peak (intact IgG )

A liquid preparation in which the high molecular weight component as measured by SE-HPLC after storage for 6 weeks at a temperature of 5 캜 3 캜 is 0 to 0.3%;

- a liquid formulation having a temperature of 5 ° C ± 3 ° C and a high molecular weight component as measured by SE-HPLC after storage for 6 weeks under confined conditions of 0 to 0.3%;

A liquid preparation in which the high molecular weight component as measured by SE-HPLC after storage for 6 weeks at a temperature of 40 ° C ± 2 ° C is 0 to 0.9%;

- a liquid formulation having a temperature of 40 ° C ± 2 ° C, a relative humidity of 75 ± 5% and a high molecular weight component as measured by SE-HPLC after storage for 6 weeks in an airtight condition of 0 to 0.9%;

- a liquid formulation having a high molecular weight component as measured by SE-HPLC of from 0 to 1.4% after storage for 3 weeks at a temperature of 45 ° C ± 2 ° C.

Main content (main peak)

A liquid preparation in which the main component is 99.7% to 100% as measured by SE-HPLC after storage for 6 weeks at a temperature of 5 ° C ± 3 ° C;

A liquid formulation having a temperature of 5 ° C ± 3 ° C and a main component of 99.7 to 100% as measured by SE-HPLC after storage for 6 weeks under confined conditions;

A liquid preparation in which the main component is 95.0% to 100% as measured by SE-HPLC after storage at 40 ° C ± 2 ° C for 6 weeks;

A liquid formulation having a temperature of 40 ° C ± 2 ° C, a relative humidity of 75 ± 5% and a main component of 95.0 to 100% as measured by SE-HPLC after storage for 6 weeks under confined conditions.

Low molecular weight components (peaks with a retention time back on the basis of the main peak (intact IgG )

- a liquid formulation with a low molecular weight component of 0.0% as measured by SE-HPLC after storage at 5 ° C ± 3 ° C for 6 weeks;

- a liquid formulation with a low molecular weight component of 0.0% as measured by SE-HPLC after storage at 5 ° C ± 3 ° C for 6 weeks under confined conditions;

- a liquid formulation in which the low molecular weight component as measured by SE-HPLC after storage for 6 weeks at a temperature of 40 ° C ± 2 ° C is 0 to 4.0%;

- a liquid formulation having a low molecular weight component as measured by SE-HPLC of 0 to 4.0% after storage for 6 weeks at a temperature of 40 ° C ± 2 ° C and a relative humidity of 75 ± 5%.

Intact immunoglobulin G content

A liquid preparation in which the content of intact immunoglobulin G (Intact IgG%) measured from non-reducing CE-SDS after storage for 6 weeks at a temperature of 5 ° C ± 3 ° C is 98.0% to 100%;

A liquid formulation at a temperature of 5 ° C ± 3 ° C and a content of intact immunoglobulin G (Intact IgG%) as measured by non-reducing CE-SDS after storage for 6 weeks in an airtight condition of 98.0% to 100%;

- a liquid preparation in which the content of intact immunoglobulin G (Intact IgG%) measured by non-reducing CE-SDS after storage for 6 weeks at 40 ° C ± 2 ° C is 93.1% to 100%;

- The content of intact immunoglobulin G (Intact IgG%), measured by non-reducing CE-SDS after storage for 6 weeks at 40 ° C ± 2 ° C, 75 ± 5% relative humidity and closed conditions, is 93.1% to 100% Liquid formulation.

Number of insoluble particle

- liquid preparations having a number of insoluble foreign particles (1.00 [mu] m, < 100.00 [mu] m) measured from the MFI after storage for 6 weeks at a temperature of 40 [deg.] C +/- 2 deg.

A liquid preparation having a temperature of 40 ° C ± 2 ° C, a relative humidity of 75 ± 5%, and a number of insoluble foreign particles (1.00 μm ≦, <100.00 μm) measured by MFI after storage for 6 weeks in an airtight condition, ;

A liquid preparation having an insoluble foreign particle (1.00 占 퐉, <100.00 占 퐉) of 0 to 5,000 measured by MFI after storage at 45 占 폚 占 2 占 폚 for 3 weeks;

[Method for producing stable liquid preparation]

The stable liquid preparations of the present invention can be prepared using known methods and are not limited to specific methods. For example, a liquid preparation can be prepared by adjusting the pH while adding an acetate buffer solution to a solution containing glycine and a surfactant, and then adding the antibody to the mixed solution.

In one embodiment of the invention, the liquid formulation may not include a lyophilization process in manufacture, or may comprise a lyophilization process.

When the lyophilization process is not included, for example, the liquid preparation of the present invention can be prepared and placed in an airtight container immediately after the treatment such as sterilization.

When a lyophilization process is included, for example, the liquid formulation of the present invention is prepared and lyophilized, or the liquid formulation of the present invention is prepared and lyophilized and stored / stored, followed by lyophilization and / or storage / The liquid formulation according to the present invention can be prepared by replenishing or replacing components that have been removed or modified. After lyophilizing only the components excluded from the components that can be removed or modified by lyophilization and / or storage / storage in the liquid preparation of the present invention, or lyophilizing and storing / storing only the components, The liquid ingredients according to the present invention can be prepared by adding the excluded components.

[Method of using stable liquid preparation]

The stable liquid formulation according to the present invention can be used to treat diseases for which the antibody is targeted, for example, a disease in which the activity of TNF-a is detrimental. Examples of diseases that are detrimental to the activity of TNFa include, but are not limited to, sepsis, autoimmune diseases, infectious diseases, grafts, malignancies, pulmonary disorders, intestinal disorders, and cardiac disorders.

In one embodiment of the invention, the disease for which the activity of TNF-a is detrimental can be selected from rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, paediatrics, psoriasis and psoriatic arthritis.

The stable liquid preparations according to the invention may be used for once, several times or for subcutaneous administration.

The total volume of the liquid formulation is in the range of 0.2 to 10.0 mL, such as 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.5, 3.5, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 7.6, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.0 mL.

The dosage and the administration time of the liquid preparation may be varied depending on the type of disease, severity and progress of the disease, response to the health and treatment of the patient, and judgment of the treating physician. It does not. For example, one or several products containing the liquid formulation may be administered in an amount of 0.1 to 10 mg / kg (e.g., 5 to 500 mg of antibody in the case of a 50 kg patient) Alternatively, different doses may be administered weekly, biweekly, every 3 weeks, monthly, 2 months, or every 3 months. In addition, the dosage and time of administration of the liquid preparation can be determined by reference to an approved medicament containing the antibody, for example, Humira.

[Treatment method and stabilization method]

The present invention also relates to a method of treating a patient suffering from a disease to which the antibody is targeted, for example, a disease in which the activity of TNF-a is detrimental, to (A) an antibody or antigen binding portion thereof; (B) acetate buffer; (C) glycine; And (D) a stable liquid formulation comprising a surfactant and not containing at least one of a sugar, a sugar alcohol, and a metal salt, such as the activity of a TNF- This provides a way to cure the harmful disease.

The present invention also provides a kit comprising (A) an antibody or antigen binding portion thereof; (B) acetate buffer; (C) glycine; And (D) preparing a stable liquid formulation comprising a surfactant and not containing at least one of a sugar, a sugar alcohol, and a metal salt, in a liquid formulation.

In one embodiment of the therapeutic method or stabilization method, (A) the antibody may comprise a monoclonal antibody.

In one embodiment of the therapeutic method or stabilization method, (A) the antibody may comprise a fully human antibody.

In one embodiment of the above method of treatment or stabilization, (A) the antibody may comprise an antibody that binds TNF- [alpha].

In one embodiment of the method of treatment or the method of stabilization, (A) the antibody may comprise one or more of infliximab, adalimumab, sertolium febulol, and golimumim.

In one embodiment of the therapeutic method or stabilization method, the antibody (A) comprises a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and an amino acid sequence of SEQ ID NO: A light chain variable region comprising a CDR3 domain; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6.

In one embodiment of the therapeutic method or stabilization method, (A) the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7; And a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

In one embodiment of the above method of treatment or stabilization, (A) the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9; And a heavy chain comprising the amino acid sequence of SEQ ID NO: 10.

In one embodiment of the therapeutic method or stabilization method, the concentration of (A) antibody may be 50 to 150 mg / mL.

In one embodiment of the method of treatment or stabilization, (B) the acetate buffer solution may comprise an acetate salt.

In one embodiment of the treatment or stabilization method, the acetic acid salt content may be between 1 and 30 mM.

In one embodiment of the above method of treatment or stabilization, the stable liquid formulation may not contain at least one of histidine, citrate, phosphate, maleate, tartrate, and succinate.

In one embodiment of the method of treatment or stabilization, the concentration of (C) glycine may be between 100 and 300 mM.

In one embodiment of the above method of treatment or stabilization, the stable liquid formulation comprises at least one of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, Tryptophan, tyrosine, and valine.

In one embodiment of the method of treatment or stabilization, (D) the surfactant may comprise polysorbate, poloxamer, or mixtures thereof.

In one embodiment of the method of treatment or stabilization, (D) the surfactant may comprise at least one of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.

In one embodiment of the method of treatment or stabilization, (D) the surfactant may comprise polysorbate 80.

In one embodiment of the treatment or stabilization method, the concentration of (D) surfactant may be 0.01 to 1% (w / v).

In one embodiment of the treatment or stabilization method, the pH of the stable liquid formulation may be between 4.5 and 5.5.

In one embodiment of the above method of treatment or stabilization, the osmolality of the stable liquid formulation may be 200 to 400 mmol / kg.

In one embodiment of the method of treatment or stabilization, the stable liquid formulation may not contain a preservative, a chelating agent or a mixture thereof.

In one embodiment of the method of treatment or stabilization, the stable liquid formulation may further comprise an aqueous carrier, an antioxidant, or a mixture of two or more thereof.

In one embodiment of the treatment or stabilization method, the viscosity of the stable liquid formulation may be between 0.5 and 5.0 cp.

In one embodiment of the method of treatment, a stable liquid formulation can be administered subcutaneously.

[product]

The present invention also relates to the stable liquid formulation; And a container for containing the stable liquid formulation in a sealed state.

The stable liquid preparation is as described above.

In one embodiment of the invention, the container can be formed from materials such as glass, polymer (plastic), metal, and the like, but is not limited thereto. In one embodiment of the invention, the container is a bottle, vial, syringe, for example a pre-fillable or pre-filled syringe, or tube, but is not limited thereto. In one embodiment of the invention, the container can be a glass or polymer vial, or a glass or polymer free-field syringe. In one embodiment of the invention, a pre-field syringe filled with the stable liquid formulation is provided.

In one embodiment of the present invention, the interior of the pre-field syringe may be coated with silicone oil. This can be excellent in terms of plunger-stopper break loose force or dynamic glide force. In one embodiment of the present invention, the silicone oil may not be coated inside the pre-field syringe. In this case, the stability of the preparation can be excellent. The container may be a single-dose or multi-dose container.

In one embodiment of the present invention, the article is an automatic syringe, and the automatic syringe may include therein a pre-field syringe filled with the stable liquid formulation. The automatic syringe may include, for example, a cylindrical housing that houses a pre-field syringe and an actuator (e.g., a spring) that initiates dosing by applying pressure to a stopper of the pre-field syringe, ), Metals, and the like. One of the known products as an automatic syringe can be used or customized to a pre-field syringe.

In one embodiment of the invention, the product may further comprise instructions for providing a method of using the stable liquid formulation, a storage method, or both. In one embodiment of the invention, the method of use comprises the treatment of a disease to which the antibody is targeted, e. G., A disease which is detrimental to the activity of TNF-a, and may include the route of administration, have.

In one embodiment of the present invention, the product may include other tools, such as needles, syringes and the like, that are necessary from a commercial and user perspective.

Hereinafter, the present invention will be described in detail by way of examples. The following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example

Regarding the antibodies used in Examples 1-2 and Comparative Examples 1-5 and 7 below, the AvialiMEMMM manufactured by Celltrion was used and with respect to the antibody used in Comparative Example 6 below, Humira® was used.

The following methods were used to measure the physical stability and chemical stability of the liquid preparations used in the following examples.

- Appearance analysis

The clarity of the preparation was visually observed.

- Turbidity

Absorbance at 350 nm was measured using a UV-Vis spectrophotometer.

- Active ingredient content

* Size exclusion The main content (%) was determined using high performance liquid chromatography (Size Exclusion HPLC).

- high molecular weight component content

Size Exclusion The content of high molecular weight components (pre-peak;%) was measured using High Performance Liquid Chromatography (Size Exclusion HPLC).

- low molecular weight component content

Size Exclusion The content of low molecular weight components (post-peak;%) was measured using high performance liquid chromatography (Size Exclusion HPLC).

- Intact immunoglobulin G content (Intact IgG%)

The content (%) of intact immunoglobulin G was measured using Non-Reduced Capillary Electrophoresis-Sodium Dodecyl Sulfate (NR CE-SDS).

- Number of insoluble particle

The number of insoluble foreign particles (1.00 占 퐉, <100 占 퐉) was measured using Micro Flow Imaging (MFI).

- Osmolality

The osmotic pressure (mmol / kg) was measured using an osmometer (VAPRO 5520).

- Viscosity

Was measured in a 500 μL syringe at 25 ° C ± 0.1 ° C using a micro-capillary flow system (apparent shear rate range: 103 to 105 s-1) equipped with a flow cell (B05 sensor type, 50 μm cell depth)

Examples 1-2 and Comparative Examples 1-12

With respect to the liquid preparations of Examples 1-2 and 1-12, each buffer was prepared for each pH, followed by addition of an amino acid or metal salt or sugar or sugar alcohol, adding the antibody thereto, adding a surfactant To prepare samples of Table 1. The specific content of each component is as shown in Table 1. The total volume was 3 mL.

division Antibody content
(mg / mL) Surfactants Amino acids or metal salts or sugars or sugar alcohols Buffer pH Example 1 100 Polysorbate 80
0.1% (w / v) Glycine 250 mM Sodium acetate 10 mM 5.2 Example 2 100 Polysorbate 80
0.1% (w / v) Glycine 280 mM Sodium acetate 10 mM 5.2 Comparative Example 1 100 Polysorbate 80
0.1% (w / v) NaCl 30 mM,
Mannitol 5% (w / v) - 5.2 Comparative Example 2 100 Polysorbate 80
0.1% (w / v) NaCl 30 mM,
Sorbitol 5% (w / v) - 5.2 Comparative Example 3 100 Polysorbate 80
0.1% (w / v) NaCl 30 mM,
Trehalose 8% (w / v) - 5.2 Comparative Example 4 100 Polysorbate 80
0.1% (w / v) NaCl 30 mM,
Sucrose 8% (w / v) - 5.2 Comparative Example 5 100 Polysorbate 80
0.1% (w / v) Lysine 250 mM Sodium acetate 10 mM 5.2 Comparative Example 6 50 Polysorbate 80
0.1% (w / v) NaCl 105 mM
Mannitol 1.2% (w / v) Sodium phosphate
14.1 mM
Sodium citrate
7.2 mM 5.2 Comparative Example 7 100 Polysorbate 80
0.1% (w / v) Glycine 310 mM Sodium acetate 10 mM 5.2 Comparative Example 8 100 Polysorbate 80
0.1% (w / v) Glycine 250 mM Sodium citrate 10 mM 5.2 Comparative Example 9 100 Polysorbate 80
0.1% (w / v) Glycine 250 mM Sodium succinate 10 mM 5.2 Comparative Example 10 100 Polysorbate 80
0.1% (w / v) Glycine 90 mM Sodium acetate 10 mM 5.2 Comparative Example 11 100 Polysorbate 80
0.1% (w / v) Glycine 250 mM Sodium acetate 30 mM 5.2 Comparative Example 12 100 Polysorbate 80
0.1% (w / v) Glycine 250 mM Sodium acetate 50 mM 5.2

The liquid formulations prepared according to Examples 1-2 and Comparative Examples 1-7 were stored for 6 weeks at a temperature of 5 ± 3 ° C, a temperature of 40 ± 2 ° C and a relative humidity of 75 ± 5%.

In addition, the liquid preparations prepared according to Example 1 and Comparative Examples 8-12 were stored at a temperature of 45 ± 2 ° C for 3 weeks.

Appearance analysis

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 Clear Clear Clear Example 2 Clear Clear Clear Comparative Example 1 Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Comparative Example 2 Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Comparative Example 3 Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Comparative Example 4 Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Very weak turbidity
(Very slightly opalescent) Comparative Example 5 Weak turbidity
(Slightly opalescent) Weak turbidity
(Slightly opalescent) Weak turbidity
(Slightly opalescent)

As shown in Table 2, the appearance of Example 1-2 was relatively transparent compared to Comparative Example 1-5, and no change in appearance was observed with time in each storage condition.

Turbidity

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 0.0845 0.0814 0.1245 Example 2 0.1086 0.0650 0.1100 Comparative Example 1 0.1120 0.1113 0.1364 Comparative Example 2 0.1160 0.1087 0.1375 Comparative Example 3 0.1115 0.1049 0.1349 Comparative Example 4 0.1135 0.1070 0.1406 Comparative Example 5 0.1920 0.1916 0.2385 Comparative Example 6 0.1230 N / A 0.1373

From Table 3, it can be seen that Example 1-2 containing acetate buffer and glycine is the most excellent in turbidity, and it is confirmed that the absorbance is kept lower than Comparative Example 1-6 even after 6 weeks at 40 ° C .

division 45 ± 2 ° C
After 0 weeks 45 ± 2 ° C
After 3 weeks Example 1 0.0718 0.1722 Comparative Example 8 0.1439 0.5250 Comparative Example 9 0.0922 0.2127 Comparative Example 10 0.0719 0.1933 Comparative Example 11 0.0937 0.2548 Comparative Example 12 0.1096 0.5384

Table 4 shows that Example 1 containing 10 mM acetate buffer and 250 mM glycine was the best in turbidity. Especially after 3 weeks at 45 ° C, the absorbance was lower than 0.1800 and lower than Comparative Example 8-12 It can be confirmed that it is maintained.

High molecular weight component content

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 0.2 0.3 0.9 Comparative Example 1 0.4 0.4 1.0 Comparative Example 2 0.3 0.4 1.0 Comparative Example 3 0.4 0.4 1.0 Comparative Example 4 0.4 0.4 1.1

From Table 5, it can be seen that Example 1 contains the least amount of high molecular weight components under all conditions. In particular, it can be seen that Example 1 contained less than 1.0% of high molecular weight components after 6 weeks at a temperature of 40 ° C.

division 45 ± 2 ° C
After 0 weeks 45 ± 2 ° C
After 3 weeks Example 1 0.1 1.3 Comparative Example 8 0.1 1.3 Comparative Example 9 0.1 1.7 Comparative Example 10 0.1 1.3 Comparative Example 11 0.1 1.6 Comparative Example 12 0.1 1.8

Referring to Table 6, it can be seen that Example 1 contained less than 1.5% of high molecular weight components after 3 weeks at a temperature of 45 ° C.

Active ingredient content

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 99.7 99.7 95.3 Comparative Example 1 99.6 99.6 94.8 Comparative Example 2 99.6 99.6 94.7 Comparative Example 3 99.6 99.6 94.7 Comparative Example 4 99.6 99.6 94.7 Comparative Example 5 99.7 99.7 94.7 Comparative Example 6 99.6 N / A 94.6

As shown in Table 7, it can be seen that the monomer content of Example 1 was higher than that of Comparative Example 1-6 at a temperature of 40 占 폚 after 6 weeks of 95.0% or more.

division 45 ± 2 ° C
After 0 weeks 45 ± 2 ° C
After 3 weeks Example 1 99.4 95.0 Comparative Example 8 99.4 95.3 Comparative Example 9 99.4 95.1 Comparative Example 10 99.4 95.1 Comparative Example 11 99.4 94.9 Comparative Example 12 99.4 94.7

As shown in Table 8, it can be seen that Example 1 has a monomer content of 95.0% or more after 3 weeks at a temperature of 45 ° C, which is higher than Comparative Example 11-12.

Low molecular weight component content

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 0.0 0.0 3.8 Comparative Example 1 0.0 0.0 4.2 Comparative Example 2 0.0 0.1 4.3 Comparative Example 3 0.0 0.1 4.3 Comparative Example 4 0.0 0.0 4.2 Comparative Example 5 0.0 0.1 4.5 Comparative Example 6 0.2 N / A 4.8

From Table 9, it can be seen that Example 1 had a low molecular weight component content of less than 4% after 6 weeks at a temperature of 40 ° C and was lower than Comparative Examples 1-6.

The content of intact immunoglobulin G (Intact IgG%)

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 98.14 98.10 93.14 Comparative Example 1 98.14 98.14 93.07 Comparative Example 2 98.27 97.92 92.73 Comparative Example 3 98.15 97.99 92.78 Comparative Example 4 98.17 97.94 92.99 Comparative Example 5 98.15 97.97 92.69 Comparative Example 6 98.15 N / A 91.25

As shown in Table 10, it can be seen that in Example 1, the content of intact immunoglobulin G was 93.10% or more after 6 weeks at a temperature of 40 ° C, which was higher than that of Comparative Example 1-6.

The number of insoluble foreign particles (1.00 占 퐉, <100.00 占 퐉)

division 5 ± 3 ° C
After 0 weeks 5 ± 3 ° C
After 6 weeks 40 ± 2 ° C
After 6 weeks Example 1 721 2881 4973 Example 2 8888 3101 7115 Comparative Example 6 41825 N / A 51914 Comparative Example 7 3319 2864 37751

As shown in Table 11, in Example 1 or 2 using glycine at 250 or 280 mM, the number of insoluble foreign particles was 10,000 or less after 6 weeks at a temperature of 40 占 폚. However, in the case of the Humira formulation of Comparative Example 6, the number of insoluble foreign particles was more than 50,000 after 6 weeks at a temperature of 40 ° C. In the case of Comparative Example 7 using glycine at 310 mM, the number of insoluble foreign particles was more than 35,000 after 6 weeks at a temperature of 40 占 폚.

division 45 ± 2 ° C
After 0 weeks 45 ± 2 ° C
After 3 weeks Example 1 2560 4778 Comparative Example 8 7146 4834586 Comparative Example 9 655 5415 Comparative Example 10 223 1615 Comparative Example 11 526 1911 Comparative Example 12 340 4332120

As shown in Table 12, in Example 1 using glycine at 250 mM, the number of insoluble foreign particles was 5,000 or less after 3 weeks at a temperature of 45 캜. However, in Comparative Example 8 and Comparative Example 12, the number of insoluble foreign particles was 4,000,000 or more after 3 weeks at a temperature of 45 ° C.

Osmotic pressure

division 45 ± 2 ° C
After 0 weeks 45 ± 2 ° C
After 3 weeks Example 1 269 304 Comparative Example 8 282 312 Comparative Example 9 275 295 Comparative Example 10 105 125 Comparative Example 11 300 335 Comparative Example 12 340 378

As shown in Table 13, the osmotic pressure of Comparative Example 10 is less than 200 mmol / kg, which is lower than that of Example 1 and Comparative Examples 8-9 and 11-12.

Viscosity

division 45 ± 2 ° C
After 0 weeks 45 ± 2 ° C
After 3 weeks Example 1 2.6 2.6

As shown in Table 14, the viscosity of Example 1 was measured to be less than 3.0 after three weeks at a temperature of 45 캜.

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Ser Ser Leu Ser Ala Ser Val Gly   1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Tyr              20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile          35 40 45 Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Ser Ser Arg Phe Ser Gly      50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro  65 70 75 80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Arg Tyr Asn Arg Ala Pro Tyr                  85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys             100 105 <210> 8 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> Antibody <400> 8 Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser   1 5 10 15 Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr Ala              20 25 30 Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser          35 40 45 Ala Ile Thr Trp Asn Ser Gly His Ile Asp Tyr Ala Asp Ser Val Glu      50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu  65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala                  85 90 95 Lys Val Ser Tyr Leu Ser Thr Ala Ser Ser Leu Asp Tyr Trp Gly Gln             100 105 110 Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 9 <211> 236 <212> PRT <213> Artificial Sequence <220> <223> Antibody <400> 9 Met Asp Phe Gln Val Gln Ile Ile Ser Phe Leu Leu Ile Ser Ala Ser   1 5 10 15 Val Ile Met Ser Arg Gly Asp Ile Gln Met Thr Gln Ser Ser Ser Ser              20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser          35 40 45 Gln Gly Ile Arg Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys      50 55 60 Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val  65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr                  85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys Gln Arg             100 105 110 Tyr Asn Arg Ala Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile         115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp     130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu                 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp             180 185 190 Ser Thr Ser Ser Ser Ser Thr Ser Ser Ser Ser Thr Leu         195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser     210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 10 <211> 470 <212> PRT <213> Artificial Sequence <220> <223> Antibody <400> 10 Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg   1 5 10 15 Val Leu Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln              20 25 30 Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ser Ser Gly Phe Thr Phe          35 40 45 Asp Asp Tyr Ala Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu      50 55 60 Glu Trp Val Ser Ala Ile Thr Trp Asn Ser Gly His Ile Asp Tyr Ala  65 70 75 80 Asp Ser Val Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn                  85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val             100 105 110 Tyr Tyr Cys Ala Lys Val Ser Tyr Leu Ser Thr Ala Ser Ser Leu Asp         115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys     130 135 140 Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 145 150 155 160 Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro                 165 170 175 Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr             180 185 190 Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val         195 200 205 Val Thr Val Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn     210 215 220 Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro 225 230 235 240 Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu                 245 250 255 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp             260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp         275 280 285 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly     290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 305 310 315 320 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp                 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro             340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu         355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn     370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr                 405 410 415 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys             420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys         435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu     450 455 460 Ser Leu Ser Pro Gly Lys 465 470

Claims (27)

(A) an antibody or antigen binding portion thereof;
(B) acetate buffer;
(C) glycine; And
(D) a surfactant,
A sugar alcohol, a sugar, a sugar alcohol, and a metal salt. The stable liquid formulation according to claim 1, wherein (A) the antibody comprises a monoclonal antibody. The stable liquid formulation of claim 1, wherein (A) the antibody comprises a fully human antibody. The stable liquid formulation of claim 1, wherein (A) the antibody comprises an antibody that binds TNF- [alpha]. The stable liquid formulation of claim 1, wherein (A) the antibody comprises at least one of infliximag, adalimumab, sertolium febulol, and golimumim. The antibody of claim 1, wherein the antibody (A) comprises a light chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: domain; And
A heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6. The antibody of claim 1, wherein the antibody (A) comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7; And a heavy chain variable region comprising the amino acid sequence of SEQ ID &lt; RTI ID = 0.0 &gt; 8. &Lt; / RTI &gt; The antibody of claim 1, wherein the antibody (A) comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9; And a heavy chain comprising the amino acid sequence of SEQ ID NO: 10. The stable liquid formulation according to claim 1, wherein (A) the concentration of the antibody is 50 to 150 mg / mL. The stable liquid formulation according to claim 1, wherein (B) the acetate buffer solution comprises an acetate salt. The stable liquid formulation according to claim 1, wherein the acetic acid salt content is 1 to 30 mM. The stable liquid formulation of claim 1, wherein the stable liquid formulation does not comprise at least one of histidine, citrate, phosphate, maleate, tartrate, and succinate. The stable liquid formulation according to claim 1, wherein (C) the concentration of glycine is 100 to 300 mM. The stable liquid formulation of claim 1, wherein the stable liquid formulation is selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, A stable liquid formulation that does not contain more than one. The stable liquid formulation of claim 1, wherein (D) the surfactant comprises polysorbate, poloxamer, or a mixture thereof. The stable liquid formulation of claim 1, wherein (D) the surfactant comprises at least one of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. The stable liquid formulation of claim 1, wherein (D) the surfactant comprises polysorbate 80. The stable liquid formulation according to claim 1, wherein (D) the concentration of the surfactant is 0.01 to 1% (w / v). The stable liquid formulation according to claim 1, wherein the pH is 4.5 to 5.5. The stable liquid formulation according to claim 1, wherein the osmolality concentration is 200 to 400 mmol / kg. The stable liquid formulation according to claim 1, which does not contain a preservative, a chelating agent or a mixture thereof. (A) 50-150 mg / mL of antibody or antigen binding portion thereof;
(B) an acetic acid salt buffer containing 1 to 30 mM acetate;
(C) glycine 100 to 300 mM; And
(D) 0.01 to 1% (w / v) of a surfactant,
A sugar alcohol, a sugar, a sugar alcohol, and a metal salt. (A) a light chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6, Binding portion 50 to 150 mg / mL;
(B) an acetic acid salt buffer containing 1 to 30 mM acetate;
(C) glycine 100 to 300 mM; And
(D) 0.01 to 1% (w / v) of a surfactant,
A sugar alcohol, a sugar, a sugar alcohol, and a metal salt. (A) a light chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 1, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 2, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3; And a heavy chain variable region comprising a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 5, and a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 6, Binding portion 100 mg / mL;
(B) Acetate buffer solution containing 10 mM acetic acid salt;
(C) glycine 250 mM; And
(D) 0.1% (w / v) of a surfactant,
A sugar alcohol, a sugar, a sugar alcohol, and a metal salt. The stable liquid formulation according to any one of claims 1 to 24, for subcutaneous administration. A pre-filled syringe filled with the stable liquid formulation according to any one of claims 1 to 24. An auto-injector wherein the pre-field syringe of claim 26 is included therein.